Category: Assay Development and Screening
The accurate, rapid and high-throughput measurement of IgG is essential in the development and manufacture of most therapeutic antibodies. Monoclonal antibodies are becoming increasingly dominant in biopharmaceuticals, where a vast number of samples must be screened for the development of each potential therapeutic.
Here we report the development of a novel, rapid, and simple fluorescence polarization based assay for high-throughput titer measurement of IgG and Fc-containing derivatives. It uses fluorescently labeled protein G that binds IgG. Upon binding, the depolarization of emitted light decreases, this can be detected to report on the presence of IgG. The CLARIOstar®, PHERAstar® and POLARstar® microplate readers exhibit excellent assay quality when used with the ValitaTITER assay, which enables a high-throughput, simple, precise method for quantification of IgG.
The combination of ValitaTITER assay with BMG LABTECH multi-mode readers allowed for quantification of IgG in the range between 2.5-80 mg/l. Comparing the microplate-based quantitation to conventional HPLC quantification revealed a high correlation of both methods with R² = 0.9926. While providing comparable results, the ValitaTITER assay for IgG quantification uses sample straight cell culture supernatant without any complex sample preparation, making the assay exceptionally fast.
Andrea Krumm– Applications Specialist, BMG LABTECH GmbH, Ortenberg, Baden-Wurttemberg, Germany
BMG LABTECH GmbH
Ortenberg, Baden-Wurttemberg, Germany
Andrea Krumm is an Applications Specialist with BMG LABTECH. She graduated in Biotechnology at the University of Applied Sciences, Darmstadt, Germany, and received her doctorate at Mainz University, Germany, in the field of cancer cell biology. Since 2016 she provides applicative support for BMG LABTECH microplate readers and informs about novel assays in application notes.